Table 2.
Fidelity of the wild-type and K1061A mutant pol ζ proteins
| dNTP·template | Wild-type | K1061A | Increaseb | ||
|---|---|---|---|---|---|
| kcat/Km (μM/min) | finca | kcat/Km (μM/min) | finca | ||
| dGTP·G | 5.2 × 10−4 | 3.1 × 10−4 | < 2 × 10−6 | < 2.5 × 10−6 | > 120 |
| dATP·G | 2.8 × 10−4 | 1.6 × 10−4 | < 2 × 10−6 | < 2.5 × 10−6 | > 64 |
| dTTP·G | 5.2 × 10−4 | 3.1 × 10−4 | < 2 × 10−6 | < 2.5 × 10−6 | > 120 |
| dCTP·G | 1.7 | NA | 0.81 | NA | NA |
| dGTP·A | 2.4 × 10−4 | 2.4 × 10−4 | < 2 × 10−6 | < 4.8 × 10−5 | > 5.0 |
| dATP·A | 1.9 × 10−4 | 1.9 × 10−4 | < 2 × 10−6 | < 4.8 × 10−5 | > 4.0 |
| dTTP·A | 1.0 | NA | 0.042 | NA | NA |
| dCTP·A | 6.9 × 10−5 | 6.9 × 10−5 | < 2 × 10−6 | < 4.8 × 10−5 | > 1.4 |
| dGTP·T | 4.6 × 10−3 | 4.2 × 10−3 | < 2 × 10−6 | < 9.5 × 10−6 | > 440 |
| dATP·T | 1.1 | NA | 0.21 | NA | NA |
| dTTP·T | 4.3 × 10−4 | 3.9 × 10−4 | < 2 × 10−6 | < 9.5 × 10−6 | > 41 |
| dCTP·T | 2.6 × 10−4 | 2.4 × 10−4 | < 2 × 10−6 | < 9.5 × 10−6 | > 25 |
| dGTP·C | 5.5 | NA | 0.40 | NA | NA |
| dATP·C | 4.5 × 10−4 | 8.2 × 10−5 | < 2 × 10−6 | < 5.0 × 10−6 | > 16 |
| dTTP·C | 1.1 × 10−3 | 2.0 × 10−4 | < 2 × 10−6 | < 5.0 × 10−6 | > 40 |
| dCTP·C | 1.2 × 10−4 | 2.2 × 10−5 | < 2 × 10−6 | < 5.0 × 10−6 | > 4.4 |
NA, not applicable.
aThe frequency of misincorporation (finc) of a given incorrect dNTP is equal to the kcat/Km for incorporating the incorrect dNTP divided by the kcat/Km for incorporating the corresponding correct dNTP.
bThe fold increase in fidelity for the K1061A mutant protein relative to the wild-type protein is equal to the finc for the wild-type protein divided by the finc for the mutant protein.